Multiple domestication events explain the origin of Gossypium hirsutum landraces in Mexico

Abstract Several Mesoamerican crops constitute wild‐to‐domesticated complexes generated by multiple initial domestication events, and continuous gene flow among crop populations and between these populations and their wild relatives. It has been suggested that the domestication of cotton (Gossypium hirsutum) started in the northwest of the Yucatán Peninsula, from where it spread to other regions inside and outside of Mexico. We tested this hypothesis by assembling chloroplast genomes of 23 wild, landraces, and breeding lines (transgene‐introgressed and conventional). The phylogenetic analysis showed that the evolutionary history of cotton in Mexico involves multiple events of introgression and genetic divergence. From this, we conclude that Mexican landraces arose from multiple wild populations. Our results also revealed that their structural and functional chloroplast organizations had been preserved. However, genetic diversity decreases as a consequence of domestication, mainly in transgene‐introgressed (TI) individuals (π = 0.00020, 0.00001, 0.00016, 0, and 0, of wild, TI‐wild, landraces, TI‐landraces, and breeding lines, respectively). We identified homologous regions that differentiate wild from domesticated plants and indicate a relationship among the samples. A decrease in genetic diversity associated with transgene introgression in cotton was identified for the first time, and our outcomes are therefore relevant to both biosecurity and agrobiodiversity conservation.


| INTRODUC TI ON
Several studies have documented multiple independent domestication events and a recurrent gene flow among crop populations and their wild relatives. These events have often occurred in Mesoamerica, a region recognized as the main center of origin of crop domestication, mainly induced by the diffusion of seeds and vegetative propagules carried out by humans (Kraft et al., 2014;Moreno-Letelier et al., 2020;Roullier et al., 2013;Zizumbo-Villarreal & Colunga-GarcíaMarín, 2010).
The cultural and biological aspects related to the use and management of plants have caused the formation of wild-to-domesticated complexes, which are formed by wild populations; landraces; breeding lines (BL); feral individuals; transgene-introgressed (TI) wild populations; TI-landraces; and genetically modified BL Velázquez-López et al., 2018; Figure 1). Wild-to-domesticated complexes have been documented in beans, maize, tomato, pumpkin, and avocado crops, in which genetic diversity is differently maintained and modified by humans while also being influenced by natural evolutionary processes (Castellanos-Morales et al., 2019;Chen et al., 2009;Moreno-Letelier et al., 2020;Motta-Aldana et al., 2010;Razifard et al., 2020). Within the wild-to-domesticated complexes, local varieties have main roles in both processes of domestication and conservation of genetic resources. The introgression of alleles from landraces to BL is often more successful than the introgression from wild populations, since the former are better adapted to agricultural conditions and have less fertility or sterility issues . Local varieties usually have more phenotypic and genetic diversity than BL as well as higher levels of gene flow with wild relatives and BL, promoted through seed exchange and/or breeding (Camacho-Villa et al., 2005;Casañas et al., 2017;Taitano et al., 2019).  Brubaker and Wendel (1994) in which the north-western Yucatán Peninsula is suggested as the primary site for the earliest stages of domestication. Yucatanese corresponds to a wild race of cotton recognized as truly wild, from which punctatum, palmeri, and latifolium emerged, which are recognized as wild or agronomically primitive forms, from which BL were subsequently obtained.
Gossypium hirsutum L. (1763) is a cotton species with high biological, economical, and cultural importance worldwide. Mexico has been recognized as its center of origin, due to the occurrence of wild populations along its coastal dunes (Pérez-Mendoza et al., 2016;Ulloa et al., 2005;Wegier et al., 2011). Besides, Mexico has been considered the most ancient domestication area of this cotton species. Archeological evidence suggests that cotton has been used for more than 4000 years in Mesoamerica, and a high phenotypic and genotypic variation has been reported in local cotton varieties (Smith & Stephens, 1971). Historical records such as codices and chronicles of the Spanish Conquest indicate that cotton was already cultivated and used throughout the current Mexican territory by pre-Hispanic cultures, with purposes such as weaving textiles and practicing rituals (FAMSI, 2021a(FAMSI, , 2021bRuiz y Sandoval, 1884;Sahagún, 1829). In the 19th and 20th centuries, intensive cotton cultivation programs were promoted in Mexico. During this period, cotton was considered the "white gold," due to its high economic relevance (Aboites, 2013).
However, the intensive production of cotton crops faced significant losses caused by pests. Genetically modified cotton began to be cultivated in 1996, looking for pest problems solutions (James, 2016).
Currently, the production of transgenic cotton has replaced the farming of conventional BL, but numerous indigenous and local communities continue cultivating native landraces within the maize polyculture system called milpa, as well as in monoculture systems.
The milpa is a traditional agricultural system, as well as a culturally important socio-ecological system in Mexico. Plant species cultivated in the milpa are selected to favor their growth among themselves and agricultural management is aimed at maintaining healthy soil (Moreno-Calles et al., 2013). Maize, beans, and squash are the most frequently cultivated crops in the milpa, but in Mexican territory, it is possible to find other species intercropped (Moreno-Calles et al., 2013;Novotny et al., 2021). In some milpas, cotton landraces are associated with corn or squash ( Figure 1). Local varieties of cotton include brown (or coyuchi), green, and white cotton, which remain important to the cultural and economic activities of indigenous peoples and local communities (Pérez-Mendoza et al., 2012).
Research regarding cotton domestication, history, and production in Mexico is extensive (Aboites, 2013;Mikulska, 2001;Ruiz y Sandoval, 1884;Sahagún, 1829). However, it is necessary to integrate information about ecological and evolutionary processes and threats to achieve cotton in situ conservation. Although wild populations and domesticated plants have different evolutionary histories, they can maintain gene flow and show introgression, since their reproductive system is compatible (Brubaker & Wendel, 1994;Velázquez-López et al., 2018). Therefore, as transgenic varieties are sown in northern Mexico, the gene flow of domesticated alleles and transgenes became a worrying issue (Wegier et al., 2011). The

Cartagena Protocol on Biosafety of the Convention on Biological
Diversity indicates that each country is responsible for regulating permits for the management and release of genetically modified organisms (GMOs). Also, for taking the necessary measures to avoid negative effects on health or biological diversity due to the use of GMOs, prioritizing the protection of species in their center of origin and genetic diversity (CBD, 2003). Mexican laws and regulations have been implemented to promote the protection of cotton genetic resources. These are the cases of the norms of the Ley de bioseguridad de organismos genéticamente modificados (Diario Oficial de la Federación, 2005). Proposals of strategies for the conservation of cotton genetic resources and scientific information, especially that providing genetic and evolutionary data, are complementary to the laws (Tobón-Niedfeldt et al., 2022). This protection category includes species or populations threatened and whose recovery and conservation need to be promoted. The main threat endangering wild populations is gene flow, and the introgression of domesticated alleles and transgenes from crops . Transgenic varieties of cotton began to be planted in Mexico in 1996, andWegier et al. (2011)  Gene flow and introgression of domesticated alleles and transgenes into wild populations can induce genetic erosion, changes in the populations' structure, phenotypic modifications, and alterations in survival rates (Jin et al., 2018;Lu, 2013). Phenotypic modifications and changes in survival rates were observed experimentally by breeding wild pumpkin (Cucurbita argyrosperma ssp. sororia) with transgenic pumpkin varieties (Cucurbita pepo Virus Resistant Transgenic). The result showed that germination and survival rates were significantly higher in the parental lines than in hybrids, while the germination time was higher and more variable in hybrids compared with parental lines (Cruz-Reyes et al., 2015).
The present study is the first report about the presence of transgenes in cotton landraces. The loss or modification of genetic diversity in landraces due to transgene introgression has been scarcely studied. However, concern about genetic erosion associated with the use of modern or transgenic cultivars has been raised indirectly, since the use of modern cultivars and the tendency to homogenize diversity in monocultures has caused disuse and reduction in the number of landraces (Guzzon et al., 2021).
Another example is bean landraces in Campeche, Mexico, one of the centers of genetic diversity of this crop species. There, Martínez-Castillo et al. (2012) observed that the reduction of local varieties caused a reduction in allelic diversity from 1979 to 2007 (diversity of Nei H = 0.18 and 0.05, respectively). Because Mexico is the center of origin, domestication, and diversity of cotton, we can raise the problem of cotton in this country as an important issue concerning biosafety. The possible loss or modification of diversity related to the presence of transgenes, coupled with the antecedents observed in bean landraces, motivated us to include landraces within the priority genetic resources that must be conserved similarly to wild populations.
Chloroplast DNA sequences have been some of the most widely used molecular markers when studying domesticated plants. Their conservative nonrecombinant nature allows reconstructing phylogenetic relationships among species while maintaining the diversity needed to distinguish different populations (Cheng et al., 2019;Dong et al., 2012;Nock et al., 2019;Tamburino et al., 2020;Yin et al., 2021). Previous studies of the structure of cotton's chloroplast genome have helped to explain the phylogeographic relationships of the Gossypium species and their evolutionary processes (Chen et al., 2017;Cheng et al., 2020;Wu et al., 2018;Xu et al., 2012).
Genus Gossypium is divided into eight diploid genome groups that are named with the letters A-G, and K, and one allopolyploid clade that refers to AD genome type (Wang, Wendel, & Jinping, 2018).
These genomic groups have been defined through taxonomic and phylogenetic analysis (Wang, Wendel, & Jinping, 2018), although deciphering which parental species was the donor of the chloroplast possessed by species of the AD genotype has been a complex task.
For this reason, it has been very useful to study the complete chloroplast genome (Li et al., 2014). Studies with the whole chloroplast genome of cotton confirmed that genome A is the chloroplast donor among parental species (Li et al., 2014). Furthermore, intraspecific genetic analyses in Mexican populations have shown that the genetic diversity of wild varieties is greater than that of domesticated populations (Wegier et al., 2011). Artificial selection and bottlenecks associated with domestication processes have commonly narrowed the genetic variation of crops (Meyer & Purugganan, 2013;Smýkal et al., 2018). This observation is important for understanding the evolution of the wild-to-domesticated cotton complex in Mexico.
Despite the importance of the chloroplast genome and its functions for plants to develop correctly, the study of diversity and structural changes in relation to the insertion of transgenes has been scarcely studied because Agrobacterium-mediated transformations in cotton are designed to modify the nuclear genome. However, due to the interaction between cytoplasmic and nuclear genomes, we cannot rule out the possible effects of transgene introgression on the chloroplast (Zhao et al., 2019). For instance, (Stegemann et al., 2003) conducted experimental crosses between transgenic tobacco varieties with chloroplast modifications (inserting the ntpII and aad genes) showing that, eventually, the transgenes are transferred to the nuclear genome, where the regulatory machinery is more effective. This study, proved that DNA escapes from the chloroplast and that its integration into the nuclear genome occurs more frequently than generally considered, therefore providing a mechanism that not only causes intraspecific but also intraorganism genetic variation (Stegemann et al., 2003).
Given that local varieties represent an important part of the genetic diversity of the wild-to-domesticated complex, they complement the current view on the evolutionary history of cotton in Mexico. Without considering the study of landraces and with a limited knowledge regarding the wild populations spread throughout the Pacific coast, it has been suggested that cotton domestication originated in the Yucatán peninsula and later spread to the rest of Mexico and other countries (Brubaker & Wendel, 1994;Grover et al., 2020;Yuan et al., 2021). Based on the analysis of the divergence between wild and domesticated populations, we examined whether cotton domestication occurred in a single or in multiple events (Figure 1g,h). Furthermore, we explored the structure and diversity of the chloroplast genome, in order to expand the understanding of the genomic effects of the use and management processes of cotton. This study aspires to provide useful information for understanding the process of domestication of cotton and bases for the conservation of valuable genetic resources.

| Sampling and data acquisition
Wild and landrace samples of G. hirsutum were collected from Nayarit, Guerrero, Oaxaca, and Yucatán during 2005, 2015, and 2019 (Table 1). Genomic DNA from 16 samples was extracted from young leaves and seeds using the CTAB protocol (Wegier et al., 2011).

| Detection of transgenes in the samples
To characterize the presence of transgenic samples among the collection, we performed a Polymerase Chain Reaction (PCR) assay.

| Chloroplast genome assembly, structure, and genetic diversity analysis
Sequencing raw data results and raw data acquired from NCBI made up a total of 954 GB of information. Each sample had 10-50× sequencing coverage and the quality of the reads was examined using FastQC v.0.11.7 (Andrews et al., 2010). We used pipeline GetOrganelle V 1.7.3.4 (Jin et al., 2020) to extract and assemble the chloroplast genome. The program performs a de novo assembly using seed and genome sequences as references. Here, we used the Ribulose 1,5-bisphosphate (RuBP) sequence from Zea mays and Gossypium hirsutum Coker 310 FR complete chloroplast genome (GenBank: NC_007944; Lee et al., 2006), as seed and reference sequences, respectively. The annotation of each genome was performed using GeSeq (Tillich et al., 2017)   sorting files and to identify genomic variants. The genomic variability was compared between samples and between the sum of genomic variants in wild, TI-wild, landrace, TI-landrace, and BL genomes. Nucleotide diversity (π), N ST index, and the Tajima's D test were calculated using DnaSPv6 (Rozas et al., 2017); and synonym and nonsynonym substitution rates (dN/dS) were calculated with PAML through PAL2NAL (Suyama et al., 2006;Xu & Yang, 2013).
The genetic diversity and selection were compared between the groups with different degrees of management, and between these and the introgressed groups. We examined population structure by performing a Bayesian spatial analysis using the package RhierBAPS (Cheng et al., 2013;Tonkin-Hill et al., 2018). The haplotype diversity was calculated using DnaSPv6 (Rozas et al., 2017). In addition, we analyzed the evolutionary history and relationships among the haplotypes and gene flow by constructing a minimum-spanning network of haplotypes using TCS in the software POPART (Leigh & Bryant, 2015).

| Phylogenetic analyses and divergence time estimation
For the phylogenetic inferences, we downloaded Gossypium's chloroplast genomes from NCBI (Table S1) and used the Theobroma cacao sequence as the outgroup. All sequences were aligned using MAFFT In order to calculate the divergence time, we took into consideration all the coding gene sequences of the assembled plastomes present in this study, and we included G. herbaceum, G. logicalyx, G. stockii, and G. thurberi as external groups. We performed two independent runs of 400,000 chain length sampling of each 1000 chains. We used the gamma site with the GTR evolutionary model and estimated the nucleotide substitution rate using JModeltest (Darriba et al., 2012). We chose a relaxed clock log-normal model together with the calibrated yule model. Three calibration points were set according to the cotton chloroplast genome type divergence: F-clade (7.23 mya), E-clade (4.28 mya), and A + AB-clade (1.53 mya) according to Chen et al's research (2016). We also used BEAUti from BEAST 2.0 (Bouckaert et al., 2019;Suchard et al., 2018) to build the xml file and performed a Convergence chain test with Tracer 1.7 . The log and tree files from both runs were concatenated using Logcombiner and the best tree was extracted with Treeannotator with 15% burnin trees, maximum clade credibility tree, and mean heights. The target tree was plotted in R (Heibl, 2008) with the phyloch and strap V 1.4 packages (Bell & Lloyd, 2014). transgenes. The sequences found in each sample are shown in Table 2.  H d = 1.00000 (4), 0.50000 (2), 0.40000 (2), 0.00000 (1), 0.60000

| Selection signature and genetic diversity of wild, landrace, and breeding lines plastomes
(2) of wild, TI-wild, landraces, TI-landraces, and breeding lines, respectively; the number of haplotypes is indicated in brackets.
In the haplotype network, it is possible to see which haplotypes are shared between the groups with different degrees of management ( Figure 4). The population structure analysis showed that the TA B L E 2 Sequences of the transgenes identified using the BLAST tools.

Sample Transgenes sequences
W_yucatan2 CP4EPSPS : AGGAC CGA GTT GGA GAT GAT CAG CTT GGC TGT TCA CCG TAA TCT GCT TCG TTT GAT TGG TTA TTG TGC TAC TTC TAA  CGA AAG ACT CCT TGT CTA CCC TTA CAT GTC TAA TGG CAGTGTTGCATCCAGGCTTAGAGGTTTGCTCAAAGAC   W_yucatan3  CP4EPSPS: AACTT GTT TGG GTC TTT GAG CAA ACC TCT AAG CCT GGA TGC AAC ACT GCC ATT AGA CAT GTA AGG GTA GAC AAG  GAG TCT TTC Table S3 determined that the genetic difference between wild and TI-wild was higher than that between wild populations and landraces.

| Evolutionary relationships of the wild-todomesticated complex of cotton and among the Gossypium species
In order to develop the tree topologies, we used SSC, LCS + IR, and the whole chloroplast dataset. In the LCS + IR tree, the lower cluster showed the well-known phylogenetic topology of Gossypium genus, clustered by genome type: A + AD, F, D, B, and C + G + K. Our   Solanum lycopersicum (Tamburino et al., 2020).
When comparing the homologous regions between genomes, we identified patterns that distinguish the wild ones from the domesticated ones, exhibiting the differences in their evolutionary path- TA B L E 4 Summary of genomic variants in plastomes of wild, landraces, and breeding lines cotton (Gossypium hirsutum).
genes could work as particular markers to distinguish domesticated from wild cotton. dhB is a transmembrane gene with translocase function involved in photosynthesis under humidity stress conditions (Horváth et al., 2000); rrn16S is a cellular component of the ribosomal subunit with high diversity, e.g., in Bulbophyllum (Tang et al., 2021); and trnG-GCC is a transfer RNA mentioned as a variable and useful gene for taxonomic identification in Acanthaceae and Balsaminaceae, it is also used to analyze the effects of domestication F I G U R E 4 Haplotype network analysis. Median-joining network based on CDS regions. Each ball represents a haplotype, the size of which is proportional to its overall frequency in the wild-to-domesticated cotton complex. The number in parentheses indicates the number of mutational changes between two different haplotypes, and black dots correspond to missing haplotypes. The Yucatán Peninsula contains the highest diversity of haplotypes within the analyzed dataset. Wild individuals analyzed present exclusive haplotypes, compared with samples from domesticated.

F I G U R E 5
Phylogenetic tree of Gossypium inferred using RAxML from SSC dataset. Colored rectangles frame the genomic clades and shading color block marked the wild-to-domesticated cotton complex, pink shows the BL, Hutchinson's landraces and TI-landraces clade, wild clade in blue, landraces clade in green, and wild+landraces in purple.

| Genes under selection through domestication
It is possible to identify different selection signals among crops and wild relatives associated with variable environments and domestication, since (1) high productivity may be related to an increase in en- The genes that showed selection signals had most of the genomic variants. Thus, they could be potential candidates for molecular markers in future research. They are involved in diverse cellular functions: rpoC2 partakes in the transcription process (Tadini et al., 2020); clpP cleaves peptides into proteins in a process that requires ATP hydrolysis (Majeran et al., 2019); accD presumably takes place in fatty acid biosynthesis (Bryant et al., 2011); cemA is involved in transmembrane proton transporter activity ; rps11 is implicated in the translation process and the viral interactions ; ycf1 and ycf2 (possible ATPasa) are genes with unknown functions in the chloroplast genome, but they are essential to protein import into chloroplast stroma (Drescher et al., 2000;Kikuchi et al., 2013). Most of these genes were under a purifying selection due to nonsynonymous mutations that could be harmful to the production of energy processes (Du et al., 2020;Wang, Zhou, et al., 2018;Xu et al., 2015). Only the ycf1 gene showed positive selection among landraces, as it has been observed in species like Citrus genus (Carbonell-Caballero et al., 2015). This could be an indicator that native varieties are preserving new alleles that allow them to survive in different environments.
F I G U R E 6 (a) Phylogenetic network constructed with the LSC + IR dataset, the outgroup species was formed with a representation of each genome type of Gossypium. (b) Relationships among our samples in four main groups. The length of the branches showed the divergence between wild and domesticated G. hirsutum. Scale marks the substitutions per site and the number of branches is bootstrap support. Shading color ovals marked the wild-to-domesticated cotton complex, pink shows the BL and TI-landraces clade, wild clade in blue, landraces clade in green, and wild+landraces in purple.

| Domestication and introgression modify the genetic diversity into wild-todomesticated cotton complex
Human selection operating through plant domestication commonly leads to a reduction in genetic diversity (Smýkal et al., 2018).
Here, wild populations contained more diversity than domesticated stands, mainly in the number of STRs (Table 4). This is consistent presence of transgenes (Cry1Ab/Ac, Cry2Ab, and CP4EPSPS) in wild and landraces can be considered as markers that exhibit the recent and continuous gene flow and introgression.
The introgression of domesticated alleles and transgenes is considered a factor that modifies the genetic diversity of the nuclear genome (Ellstrand et al., 2002;Rojas-Barrera et al., 2019;. Interestingly, plastomes of transgenic samples showed less diversity and according to RhierBAPS, haplotype network, and phylogenetic analysis (Figures 4 and 5) landraces plastomes shared ancestor with the BL, which is not the case in wild plastomes with transgenes. The introgression of transgenes in the wild-todomesticated cotton complex could succeed via pollen and through the transgenic seeds' establishment and subsequent pollination. But the possible effect on diversity will depend on whether the donor of the chloroplast genome to the next generation is transgenic or nontransgenic as described in Figure S4.
The study of the diversity of plastomes through the introgression of domesticated alleles or transgenes is scarce because it is a nonrecombinant genome and the Agrobacterium-mediated transformation through which transgenes are inserted is directed to the nuclear genome (Tzfira & Citovsky, 2006). However, nuclear and cytoplasmic genomes are highly dynamic, and the transcriptional activity of chloroplast requires coordination with the nuclear gene expression, for instance, the interplay of plastid-encoded polymerases and nuclearencoded polymerases is necessary for the correct development of functional chloroplast (Tadini et al., 2020). Therefore, nuclear or

| Multiple events of domestication generated cotton landraces in Mexico
Various molecular markers have been used to elucidate Gossypium's evolutionary history and chloroplast has proven to be the most reliable for resolving its uncertainties, due to its maternal inheritance (Chen et al., 2016(Chen et al., , 2017Wu et al., 2018). However, the presence of two structural haplotypes with respect to the SSC region, added to include high variability regions, can generate discrepancies in the topologies (Figure 7 and Figure S3). Based on the less variable regions, we reconstructed a phylogeny similar to the previously reported ones, in which G. herbaceum, G. arboreum, and G. thurberi are the closest taxa to the AD clade (Figure 7; Wu et al., 2018;Xu et al., 2012). In addition, the phylogenetic analysis supported the identity and occurrence of the wild populations described by Wegier et al. (2011) and Alavez et al. (2021).
The domestication of G. hirsutum originated in Mesoamerica.
More exactly, the hypothesis proposed by Brubaker and Wendel (1994) suggested that domestication began through the selection of wild populations from the Yucatán peninsula. In order to do this, they took into account Hutchinson (1951), Fryxell (1979), and Stephens (1958) classifications, in which yucatanese corresponds to a wild race of cotton considered truly wild. It was from this taxon, according to these authors, that the punctatum, palmeri, and latifolium varieties emerged, which are recognized as wild or agronomically primitive forms, from which BL was subsequently obtained (Figure 1h). Additionally, the results provided evidence that genetic resources from the Yucatán Peninsula metapopulation (including the Guatemalan distribution area) were used in the development of BL, and that landraces played an important role in the domestication process. Unlike trees, the phylogenetic network suggested more than one evolutionary history. Although the divergence between wild and BL was lower than BL and landraces, alternate branches did not reject any alternative topology. The position of W_banderasbay and L_oaxbrown as the basal node in the tree shown in Figure 7 and the divergence observed in the network indicated multiple domestication events. It has been observed that the Banderas Bay holds higher haplotype and morphological diversity (e.g., form of leaf, fiber characteristics, and foliar epidermal characters) than the rest of metapopulations (Uscanga, 2013;Vega, 2015;Wegier et al., 2011).  (Smith & Stephens, 1971); and in the Balsas region (dated in 1200 years BP). The divergence time estimated from coding regions of chloroplast suggested that the domestication could have started 11,700 years ago (Bjerregaard & Peters, 2017). However, there is a variation range in the node calibration shown with the blue bar ( Figure 6).

| Implication to conservation of wild population and landraces in Mexico
The Using only chloroplast limits the differentiation between introgression events that occurred in the past from recent events, therefore we consider that further research should include the chloroplast of the rest of metapopulation, more samples of landraces; and should incorporate nuclear genome markers. The aforementioned actions will allow detailing evolutionary history; recognizing the origin of the genetic material used in crops and research throughout the world; and providing information to describe the transgenes dispersion routes . This information is key for sharing the benefits and avoiding harm to the local and indigenous communities, which are fundamental to the domestication processes and to the protection of genetic resources.

DATA AVA I L A B I L I T Y S TAT E M E N T
Assemblies sequences were deposited to NCBI. Vcf files associated are available in: https://www.resea rchga te.net/publi catio n/35897 7040_chlor oplas t-vcftar All code used to perform our analyses and figures are available: https://github.com/conse rvati ongen etics/ Multi ple_domes ticat ion_events_expla in_the_origin_of_landr aces_of_Gossy pium_hirsu tum_in_Mexico.git

B EN EFIT S H A R I N G
The benefits of this research include the generation of information on local varieties requested by local communities. The results were communicated to collaborators in the local communities, and their comments have been included in the discussion. In addition, they are recognized in the document as authors or in the acknowledgments according to their personal decision.